Abstract
Dengue viruses (DENV) cause the most common arboviral disease afflicting men. Clinical manifestations range from asymptomatic to dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS). The mechanisms involved in the disease pathogenesis are not fully understood. The severity of the disease seems to be influenced by both viral and host factors. Subgenomic replicons of DENV can be used to study viral replication mechanisms and evaluate the effects of antiviral drugs on viral replication. The objective was to generate and characterize biologically a replicon from a clinical isolate of DENV-3, as part of our studies to understand how this new isolate interacts with cells. To obtain this replicon several RT-PCR fragments encoding the non-structural proteins genes were cloned in high-copy vectors, and used to assemble the replicon in a BAC plasmid vector containing a synthetic DNA molecule encoding the 5′ and 3′ ends of a viral cDNA with a T7 DNA-dependent RNA polymerase promoter and a ribozyme. In vitro transcribed RNA recovered from this BAC plasmid was transfected into C6/36 mosquito cells, and dengue virus protein expression was assessed by indirect immunofluorescence using polyclonal antibodies. The results showed that the replicon was replicated efficiently in cells, demonstrating successful assembly of a DENV-3 replicon.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
